US10359388B2 - Arrangement and method for the electrochemical analysis of liquid samples by means of lateral flow assays - Google Patents

Arrangement and method for the electrochemical analysis of liquid samples by means of lateral flow assays Download PDF

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US10359388B2
US10359388B2 US14/377,765 US201314377765A US10359388B2 US 10359388 B2 US10359388 B2 US 10359388B2 US 201314377765 A US201314377765 A US 201314377765A US 10359388 B2 US10359388 B2 US 10359388B2
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carrier
arrangement
front side
electrode
membrane
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US20150008144A1 (en
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Walter Gumbrecht
Peter Paulicka
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Siemens AG
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Siemens AG
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
    • G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/40—Semi-permeable membranes or partitions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/48707—Physical analysis of biological material of liquid biological material by electrical means

Definitions

  • the present embodiments relate to an arrangement and a method for electrical detection of liquid samples using lateral flow assays.
  • Lateral flow assays are in widespread use in in-vitro diagnostics (IVD). Lateral flow assays are simple in terms of handling and very cost-effective. Disadvantages of lateral flow assays include a low sensitivity, a low multiplexity and a poor quantifiability of the results.
  • a good quantifiability may be achieved by optical, magnetic and electrical methods, but heretofore with very low multiplexity (e.g., simultaneous measurement at a plurality of spatially separate measurement points).
  • U.S. Pat. No. 6,896,778 discloses an arrangement in which, for a good multiplexity, gold electrodes are arranged above cutouts of an electrically insulating carrier as an array, and the cutouts are filled with membranes composed of a polymer/microfiber matrix material.
  • the membranes are spatially separated from one another.
  • the membranes are ion-selective, and not suitable, for example, for immunosensors in immunoassays.
  • the capture antibodies are to be immobilized directly on the gold electrodes or sensors.
  • a carrier including insulator layers and gold electrodes that is described in U.S. Pat. No. 6,896,778, where the gold electrodes are arranged above cutouts in the insulator layers in array form, a small cavity in each case arises above the gold electrodes through the surrounding insulator layer.
  • liquid is applied directly or via a lateral flow paper as a membrane above the arrangement, air bubbles arise in the region of the cavities and during a measurement, lead to a failure of the respective electrodes with air inclusions.
  • the present embodiments may obviate one or more of the drawbacks or limitations in the related art.
  • an arrangement and a method for the electrical detection of liquid samples that enable a good multiplexity in conjunction with very good sensitivity and quantifiability are provided.
  • good multiplexity may be a multiplexity (e.g., in the range of 3- to 10-plex (sensors)).
  • an arrangement and a method that enable a reliable measurement e.g., without disturbing air inclusions above the electrodes) are provided.
  • the arrangement for the electrical detection of liquid samples by lateral flow assays includes a membrane arranged on a front side of a first carrier.
  • the first carrier is embodied in an electrically insulating fashion, and electrically conductive electrodes are formed on the first carrier.
  • the electrodes on the front side of the first carrier are arranged between the first carrier and the membrane, in direct contact with the membrane.
  • the electrodes may form a direct contact with the membrane.
  • the contact area is maximized with the membrane arranged flat on the electrode, and when the liquid sample to be analyzed is applied to the membrane, the electrodes are in direct contact with the liquid sample. Air bubbles or air inclusions that may impede or completely prevent a measurement are prevented by the direct contact of electrodes and membrane, and thus also the direct contact of electrodes and the liquid sample.
  • a reliable measurement of the sample including in the case of multiplex measurement (e.g., with a plurality of sensors simultaneously), with very good sensitivity and quantifiability of the sample is provided.
  • the membrane may be configured as a closed layer via which the electrodes (e.g., all the electrodes) are connected to one another. This enables a lateral liquid transport (e.g., lateral flow) completely via the membrane (e.g., via all the electrodes).
  • the membrane may include or be a lateral flow paper (e.g., composed of nitrocellulose). Lateral flow paper has a high porosity and absorbs the liquid sample well and transports the liquid sample well to the electrodes. This leads there to a good wetting of the electrodes with the sample liquid to be examined. A good electrical contact via membrane saturated with liquid sample between electrodes is thus made possible. Nitrocellulose is cost-effective and, used as a membrane, has the properties described above.
  • the electrodes may be metal electrodes (e.g., composed of gold). Electrodes composed of metal may be relatively stable, and gold electrodes, for example, may be used well electrochemically since the gold electrodes may lead to temporally stable measurement signals and chemically are substantially inert.
  • the electrodes may be electrically contact-connected on the front side of the carrier (e.g., in an edge region in which the membrane is not arranged). This affords advantages, for example, if the rear side may not readily be reached for electrical contacts (e.g., as a result of encapsulation).
  • the carrier may also have in each case in the region of a respective electrode an opening passing through its thickness, from the front side to the rear side.
  • the electrode is electrically contact-connected conductively from the front side to the rear side of the carrier. This makes it possible to prevent electrical short circuits between electrode contacts (e.g., upon contact with sample liquid on the front side of the carrier).
  • the carrier may include a plurality of insulating layers (e.g., layers composed of polymers and/or layers that are connected to one another by lamination).
  • Laminated carriers composed of polymers are printed circuit boards, for example, that may be produced cost-effectively.
  • the membrane may be arranged in a sandwich-like fashion firstly between the front side of the first carrier in direct contact with the electrically conductive electrodes (e.g., working electrodes) of the first carrier and secondly a rear side of a second carrier in direct contact with at least one electrode (e.g., counterelectrode; exactly one electrode on the rear side of the second carrier).
  • This arrangement enables a compact construction and short paths via the membrane between electrodes in order to establish a voltage between the working electrodes and the counterelectrode.
  • Each electrode on the front side of the first carrier may be in each case electrically connected to the at least one electrode on the rear side of the second carrier (e.g., via an electrical measuring instrument or measuring device for measuring current and/or voltage and/or capacitance).
  • the electrodes may be arranged on the front side of the first carrier in a series in tandem or in array form.
  • the method according to one or more of the present embodiments for the electrical detection of liquid samples is effected by an arrangement described above.
  • the liquid sample is applied to the membrane and is moved by capillary forces (e.g., via the membrane to the electrodes).
  • the membrane interconnects the electrodes (e.g., electrochemically if the membrane is filled with liquid).
  • the electrodes e.g., electrochemically if the membrane is filled with liquid.
  • exactly one membrane interconnects all the electrodes. As a result, a good conductivity is provided in the case of conductive liquid between the electrodes via the one membrane.
  • the arrangement brings about a spatial and/or temporal separation of substances in the liquid sample analogously to chromatography or with different capture molecules immobilized at different locations according to the lateral flow method.
  • the spatial and/or temporal separation may be measured electrochemically by the electrodes in the form of current and/or voltage and/or charge changes.
  • the membrane may be in direct contact with the electrodes.
  • the contact area of each electrode with the membrane may thus be completely wetted with the liquid sample (e.g., without air inclusions above the electrode). This provides a good electrochemical measurement using the electrode, which would be prevented or at least impeded by, for example, air bubbles directly above the electrode.
  • the liquid sample may be a biochemical sample (e.g., a body fluid).
  • a biochemical sample e.g., a body fluid
  • urine, blood, or the information thereof, for example, may be examined.
  • FIG. 1 illustrates a schematic sectional illustration through an arrangement for electrical detection of liquid samples according to the prior art
  • FIG. 2 illustrates a schematic sectional illustration through one embodiment of an arrangement for electrical detection of liquid samples
  • FIG. 3 illustrates a schematic sectional illustration through one embodiment of the arrangement of FIG. 2 ;
  • FIG. 4 illustrates a schematic illustration in a plan view of the arrangement shown in FIG. 3 ;
  • FIG. 5 illustrates a schematic sectional illustration through one embodiment of the arrangement shown in FIG. 3 .
  • the arrangement 1 is shown schematically in sectional illustration.
  • the electrode 3 is arranged below a first carrier 2 , which is constructed, for example, from laminated polymer layers analogously to a printed circuit board.
  • the electrode 3 is laminated, for example, as a gold layer onto the carrier 2 .
  • the electrode 3 may also be, for example, adhesively bonded or applied electrolytically.
  • a cutout passing through the carrier 2 is introduced in the carrier 2 , above the electrode 3 .
  • the cutout may be configured, for example, in the form of a drilled hole or milled hole (e.g., in a circular fashion).
  • a membrane 4 Arranged in the cutout, in contact with the electrode 3 , is a membrane 4 as an ion-selective layer that completely covers the free area of the electrode 3 in the cutout.
  • the electrode 3 is electrically contact-connected via an electrical contact 6 from the rear side (e.g., from the side of the electrode 3 that is opposite relative to the membrane 4 and is not covered by the membrane 4 ).
  • a liquid sample 5 is guided via the membrane 4 and that side of the carrier 2 on which no electrodes 3 are arranged and which is opposite relative to the side with the electrodes on the carrier 2 .
  • the liquid sample is electrochemically in contact with the electrode 3 via the membrane 4 . This provides that ions may move through the membrane 4 from the liquid samples 5 to the electrode 3 .
  • a counterelectrode is in electrical contact with the liquid sample 5 .
  • a current, voltage and/or charge change at the electrode 3 may be measured by a measuring device 10 (not shown).
  • the measurement is carried out depending on the liquid sample 5 and serves for analyzing the sample 5 .
  • the measurement may be carried out depending on the sample flow rate and/or time and provides information about the composition or chemical/biochemical constituents of the sample 5 .
  • blood, urine or other body fluids may serve as samples 5 .
  • other liquids through to gases may also be examined.
  • the arrangement 1 shown in FIG. 1 may be used in a sensor array or some other arrangement of sensors (e.g., in series).
  • the electrodes 3 constitute the sensors. For a reliable measurement, it is important that no liquid 5 passes to that side of the first carrier 2 with the electrodes 3 on which the electrical contacts 6 are secured.
  • the ion-selective membrane 4 is not suitable for immunosensors, for example.
  • the complicated construction is costly to produce and difficult to handle.
  • FIG. 2 illustrates a schematic sectional illustration through an arrangement 1 according to one or more of the present embodiments for the electrical detection of liquid samples 5 .
  • An electrode 3 is arranged on a first, laminated carrier 2 , analogously to the arrangement 1 described above under the prior art.
  • the membrane 4 is arranged on the front side of the electrode 3 , which according to one or more of the present embodiments is opposite relative to the side of the electrode 3 in contact with the carrier 2 .
  • the membrane 4 is lateral flow paper, for example, that lies flat on the electrode.
  • the electrode 3 is electrically contact-connected via an electrical contact 6 on the rear side of the electrode 3 .
  • the electrode 3 In the laminated carrier 2 , in the region of the electrode 3 , on the rear side thereof, there is a cutout through which the electrical contact 6 projects as far as the electrode 3 .
  • the electrode 3 may also be contact-connected from the front side.
  • FIG. 3 and FIG. 4 show an exemplary embodiment of the arrangement 1 according to one or more of the present embodiments in which an electrical contact 6 of the electrodes 3 may be effected laterally on the front side.
  • FIG. 3 illustrates a schematic sectional illustration through an arrangement 1 analogous to the arrangement shown in FIG. 2 , but with a plurality of the electrodes 3 shown in FIG. 2 arranged in series in tandem.
  • the electrodes 3 may also be arranged as an array (e.g., in a matrix-type fashion on the first carrier 2 ).
  • FIG. 4 illustrates a plan view of the arrangement 1 shown in FIG. 3 with electrodes 3 in a series 7 .
  • a membrane 4 e.g., composed of lateral flow paper
  • electrical contacts 6 may be fitted in each case to each electrode 3 , this not being illustrated in the figures for the sake of simplicity.
  • FIG. 5 shows a construction for an electrical measurement with the arrangement 1 according to one or more of the present embodiments, as shown in FIGS. 3 and 4 , in a schematic sectional illustration.
  • a second carrier 8 with an electrode 9 is arranged on the membrane 4 .
  • the membrane 4 bears in direct contact flat against the electrode 9 .
  • the electrode 9 serves as a counterelectrode.
  • a series of electrodes 3 serving as working electrodes is arranged in direct contact with the membrane 4 .
  • the electrodes 3 are arranged on a carrier 2 , as already shown in FIGS. 2 to 4 . Consequently, the membrane 4 lies in a sandwich-like fashion between the electrodes 3 on the first carrier 2 and the electrode 9 applied in a planar fashion on the second carrier 8 .
  • the electrode 9 may be electrically connected to each electrode 3 in each case via an electrical measuring instrument or a measuring device 10 .
  • the electrodes 3 , 9 may be electrically contact-connected from the side with the carrier 2 , 8 through openings in the carrier 2 , 8 , as shown in FIG. 2 , or, as described for FIG. 4 , from that side of the electrodes 3 , 9 that is in direct contact with the membrane 4 , but in a region of the electrodes 3 , 7 in which the membrane 4 is not arranged.
  • a liquid sample 5 e.g., blood or urine
  • the membrane 4 e.g., lateral flow paper made of nitrocellulose in strip form or in some other form
  • the sample 5 is moved into and through the membrane 4 (e.g., through the porous structure of the membrane 4 ).
  • the membrane 4 is thus “filled” with the sample 5 .
  • the electrodes 3 , 9 in direct contact with the membrane 4 are electrically or electrochemically connected to one another, and a closed electric circuit is provided in each case via the measuring device 10 between the respective electrode 3 as working electrode and the electrode 9 as counterelectrode.
  • the composition of the liquid sample may be examined in a spatially and temporally resolved manner.
  • electrochemical measurements via current, voltage and/or capacitance measurements may provide information about the sample 5 situated at the location.
  • the sample composition may be analyzed, or capture molecules may be immobilized on the respective electrode 3 (e.g., different capture molecules on different electrodes 3 ) and enable the detection of individual substances in the sample 5 .
  • the capture molecules may also be immobilized in a spatially distributed manner in the membrane 4 in the region above the electrode 3 .
  • the arrangement 1 according to one or more of the present embodiments may be used in immunoassays.
  • the electrodes 3 , 9 may be used, as illustrated in the figures (see FIG. 5 ), in a measurement set-up including working electrode 3 and counterelectrode 9 .
  • at least one reference electrode RE may also be used.
  • Metal layers e.g., composed of gold or platinum
  • electrodes may be used, for example, as reference electrode.
  • Other measurement set-ups that are customary in electrochemistry may also be provided.
  • Electrodes 3 , 9 may be applied in planar fashion on the carrier 2 , 8 or in a spatially structured fashion, in series in tandem, in array form in an n ⁇ m matrix including n lines and m columns, or may be provided with different height profiles.

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US14/377,765 2012-02-08 2013-01-21 Arrangement and method for the electrochemical analysis of liquid samples by means of lateral flow assays Active 2034-10-16 US10359388B2 (en)

Applications Claiming Priority (4)

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DE102012201843.3 2012-02-08
DE102012201843A DE102012201843A1 (de) 2012-02-08 2012-02-08 Anordnung und Verfahren zur elektrischen Detektion von flüssigen Proben mit Lateral Flow Assays
DE102012201843 2012-02-08
PCT/EP2013/051026 WO2013117413A1 (de) 2012-02-08 2013-01-21 Anordnung und verfahren zur elektrochemischen analyse von flüssigen proben mit lateral flow assays

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US10204166B2 (en) * 2016-09-09 2019-02-12 Facebook, Inc. Ranking content items based on session information
WO2018148236A1 (en) * 2017-02-07 2018-08-16 Arizona Board Of Regents On Behalf Of Arizona State University Electrochemical osmolarity or osmolality sensor for clinical assessment
CN110923344B (zh) * 2019-12-19 2023-06-27 武汉中帜生物科技股份有限公司 金黄色葡萄球菌及耐甲氧西林金黄色葡萄球菌耐药基因mecA检测试剂盒及其应用
JP7398796B2 (ja) * 2020-02-27 2023-12-15 国立大学法人 東京大学 特定成分検出方法および判定方法並びにこれらの方法を用いた装置

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JP5963887B2 (ja) 2016-08-03
DE102012201843A1 (de) 2013-08-08
US20190271657A1 (en) 2019-09-05
EP2798348B1 (de) 2017-08-02
EP2798348A1 (de) 2014-11-05
US20150008144A1 (en) 2015-01-08
US10514353B2 (en) 2019-12-24
WO2013117413A1 (de) 2013-08-15
CN104136921A (zh) 2014-11-05

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